Supplemental information Nrf1 is endowed with a dominant tumor-repressing effect onto the Wnt/-Catenin -dependent and -independent signaling networks in the human liver cancer Jiayu Chen1,2,§, Meng Wang1,§, Xufang Ru1,3,§, Yuancai Xiang1,4,§, Yonggang Ren1,5,§, Xiping Liu2, Lu Qiu1,6, and Yiguo Zhang1* 1The Laboratory of Cell Biochemistry and Topogenetic Regulation, College of Bioengineering and Faculty of Sciences, Chongqing University, No. 174 Shazheng Street, Shapingba District, Chongqing 400044, China. 2Department of Biochemistry and Molecular Biology, Zunyi Medical University, No. 6 Xuefu-Xi Road, Xinpu New District, Zunyi 563000, Guizhou, China. 3Department of Neurosurgery, Southwest Hospital, Army (Third Military) Medical University, No. 29 Gaotanyan Street, Shapingba District, Chongqing 400038, China. 4Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Southwest Medical University, No. 1 at the First Section of Xianglin Road, Longmatan District, Luzhou 646000, Sichuan, China. 5Department of Biochemistry, North Sichuan Medical College, No. 55 Dongshun Road, Gaoping District, Nanchong 637000, Sichuan, China. 6School of Life Sciences, Zhengzhou University, No. 100 Kexue Avenue, Zhengzhou 450001, Henan, China. §Contributed equally to this work. *Correspondence should be addressed to Yiguo Zhang (Email:
[email protected], or
[email protected]) 1. Supplemental materials and methods 1.1 The TOPflash Luciferase reporter assay to measure β-catenin/Tcf-driven transcriptional activity. Human 293T cells (2×104) were seeded in each well of a 48-vial plate and allowed for growth to 70% confluence. The cells were co-transfected with 100 ng of the firefly luciferase reporter called TOPflash (driven by the consensus β-catenin/Tcf4-binding site) or its mutant control plasmid called FOPflash, together with 5 ng of Renilla luciferase reporter (pRL-CMV), plus 10 pmol of indicated small interference RNA targeting for Nrf1 (i.e., siNrf1) or a scrambled negative control RNA (i.e.